Engineered CRISPR/Cas9 enzymes improve discrimination by slowing DNA cleavage to allow release of off-target DNA

Engineered high-fidelity Cas9s have increased discrimination against off-targets. Kinetic analyses of Cas9-HF1 and HypaCas9 engineered Cas9 variants show that their DNA cleavage is impaired by more than 100- fold, which leads to release rather than cleavage of a bound off-target substrate.

Bibliographic Details
Main Authors: Mu-Sen Liu, Shanzhong Gong, Helen-Hong Yu, Kyungseok Jung, Kenneth A. Johnson, David W. Taylor
Format: Article
Language:English
Published: Nature Portfolio 2020-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-020-17411-1
Description
Summary:Engineered high-fidelity Cas9s have increased discrimination against off-targets. Kinetic analyses of Cas9-HF1 and HypaCas9 engineered Cas9 variants show that their DNA cleavage is impaired by more than 100- fold, which leads to release rather than cleavage of a bound off-target substrate.
ISSN:2041-1723